Deshmukh, A and Gopal, B (2020) Structural insights into the catalytic mechanism of Bacillus subtilis BacF. In: Acta Crystallographica Section F: Structural Biology Communications, 76 . pp. 145-151.
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Abstract
The nonribosomal biosynthesis of the dipeptide antibiotic bacilysin is achieved by the concerted action of multiple enzymes in the Bacillus subtilis bac operon. BacF (YwfG), encoded by the bacF gene, is a fold type I pyridoxal 5-phosphate (PLP)-dependent stereospecific transaminase. Activity assays with l-phenyl-alanine and 4-hydroxyphenylpyruvic acid (4HPP), a chemical analogue of tetrahydrohydroxyphenylpyruvic acid (H4HPP), revealed stereospecific substrate preferences, a finding that was consistent with previous reports on the role of this enzyme in bacilysin synthesis. The crystal structure of this dimeric enzyme was determined in its apo form as well as in substrate-bound and product-bound conformations. Two ligand-bound structures were determined by soaking BacF crystals with substrates (l-phenylalanine and 4-hydroxyphenylpyruvate). These structures reveal multiple catalytic steps: the internal aldimine with PLP and two external aldimine conformations that show the rearrangement of the external aldimine to generate product (l-tyrosine). Together, these structural snapshots provide an insight into the catalytic mechanism of this transaminase. © 2020.
| Item Type: | Journal Article |
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| Publication: | Acta Crystallographica Section F: Structural Biology Communications |
| Publisher: | International Union of Crystallography |
| Additional Information: | The copyright for this article belongs to the Authors. |
| Keywords: | amino acid sequence; Bacillus subtilis; catalysis; chemistry; crystallization; genetics; human; isolation and purification; procedures; protein secondary structure, Amino Acid Sequence; Bacillus subtilis; Catalysis; Crystallization; Humans; Protein Structure, Secondary |
| Department/Centre: | Division of Biological Sciences > Molecular Biophysics Unit |
| Date Deposited: | 24 Jan 2023 11:00 |
| Last Modified: | 24 Jan 2023 11:00 |
| URI: | https://eprints.iisc.ac.in/id/eprint/79417 |
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